Taking a Look at Botulism

604 Words Feb 17th, 2018 2 Pages
There are seven serotypes BoNTs of which only A, B and E are responsible for human botulism. These toxins bind to presynaptic nerve terminal of neuromuscular junction thus disrupting cellular communication resulting in muscular weakness and paralysis. Various ways were discovered for treating botulism which involve producing murine monoclonal antibodies against BoNTs. But if such antibodies are injected into human beings such antibodies would be detected as foreign entities and antibodies would be produced against monoclonal antibodies. The drawbacks of use monoclonal antibodies lead to development of antigen specific antibodies.
For generating antigen specific antibodies, human peripheral blood mononuclear cells (PBMC) were isolated from three healthy volunteers. PBMC were then immunized with interleukins and BoNT. Invitro immunized cells were then isolated and RNA was extracted from it. The RNA was then reverse transcribed to form cDNA. The cDNA was then used for template for production of human IgG Fd (consisting heavy chain variable and constant region) and light chain fragments using specific primers. The DNA obtained by amplifying IgG Fd and light chain was digested with respective digestion enzymes and ligated into different lambda phage and cloned. The cloned lamda phage DNA was then separated by 0.8% Agarose Gel Electrophoresis. The purified gel fragments of…

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